Enhanced T cell proliferative response to type II collagen and synthetic peptide CII (255-274) in patients with rheumatoid arthritis

Augmented production of chemokines by the interaction of type II collagen–reactive T cells with rheumatoid synovial fibroblasts

“…Previous studies on CII have revealed that it may act as a self antigen that can activate lymphocytes for the initiation of the autoimmune process in RA (15)(16)(17)(18)(19)(20). However, the effector function of CII-reactive T cells in RA remains unclear.…”

“…Lyophilized bovine CII, a generous gift from Dr. Andrew H. Kang (University of Tennessee, Memphis), was dissolved in 0.1N acetic acid at 1 mg/ml, dialyzed against 50 mM Tris, 0.2M NaCl, and then sterilized by filtering through a 0.2-m micropore filter. We used bovine CII as an antigen because the T cell response to bovine CII was strongly correlated with that of human CII and its immunodominant peptide CII (peptide 255-274) in our previous study (19). Forty micrograms per well of ovalbumin and 1 g/well of phytohemagglutinin (PHA; Gibco BRL, Grand Island, NY) were used as the negative and positive controls, respectively.…”

“…T cells (1 ϫ 10 5 /ml) and irradiated non-T cells (1 ϫ 10 5 /ml) were cocultured with or without various antigens in 96-well plates. The optimal concentration of CII was 40 g/well, and the optimal culture time was 5 days, as previously described (19).…”

“…The data are represented as a stimulation index (SI), calculated from the ratio of the counts per minute in the presence of CII to that in the absence of CII. T cell proliferative responses were considered positive if the SI was Ն2 and if the increase in cpm was Ն1,000 (19).…”

“…CII-reactive CD4ϩ T cell lines have been reported to transfer disease to naive mice (17,18). In our previous studies, we demonstrated that T cell responses to bovine CII, synthetic immunodominant peptide (peptide 255-274), and human CII are enhanced in RA patients, particularly in the joints (19). We also reported that CII-reactive T cells could induce the Th1 shift of the cytokine balance in RA (20).…”